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Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models
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Genetics
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JoVE Journal Genetics
Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models

Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models

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9,243 Views

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13:47 min

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March 29, 2019

DOI:

10.3791/59241-v

DOI:
10.3791/59241-v

•

13:47 min
March 29, 2019

•

6 Views
, , , , , , , ,
1Department of Neurology,Duke University Medical Center, 2Center for Genomic and Computational Biology,Duke University Medical Center, 3Viral Vector Core,Duke University Medical Center, 4Division of Gynecologic Oncology, Department of Obstetrics and Gynecology,Duke University Medical Center

Summary

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Targeted DNA epigenome editing represents a powerful therapeutic approach. This protocol describes the production, purification, and concentration of all-in-one lentiviral vectors harboring the CRISPR-dCas9-DNMT3A transgene for epigenome-editing applications in human induced pluripotent stem cell (hiPSC)-derived neurons.

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Lentiviral VectorEpigenome EditingHuman Induced Pluripotent Stem CellsDisease ModelsCRISPR/CasAlpha-synucleinGene ExpressionGuide RNAMolecular CloningPlasmid ConstructionNeural Progenitor CellsDNA Methyltransferase (DNMT3A)

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Tagliafierro, L., Ilich, E., Moncalvo, M., Gu, J., Sriskanda, A., Grenier, C., Murphy, S. K., Chiba-Falek, O., Kantor, B. Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models. J. Vis. Exp. (145), e59241, doi:10.3791/59241 (2019).

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